Sequence analysis of dolphin ferritin H and L subunits and possible iron-dependent translational control of dolphin ferritin gene.
BACKGROUND
Iron-storage protein, ferritin plays a central role in iron metabolism. Ferritin has dual function to store iron and segregate iron for protection of iron-catalyzed reactive oxygen species. Tissue ferritin is composed of two kinds of subunits (H: heavy chain or heart-type subunit; L: light chain or liver-type subunit). Ferritin gene expression is controlled at translational level in iron-dependent manner or at transcriptional level in iron-independent manner. However, sequencing analysis of marine mammalian ferritin subunits has not yet been performed fully. The purpose of this study is to reveal cDNA-derived amino acid sequences of cetacean ferritin H and L subunits, and demonstrate the possibility of expression of these subunits, especially H subunit, by iron.


METHODS
Sequence analyses of cetacean ferritin H and L subunits were performed by direct sequencing of polymerase chain reaction (PCR) fragments from cDNAs generated via reverse transcription-PCR of leukocyte total RNA prepared from blood samples of six different dolphin species (Pseudorca crassidens, Lagenorhynchus obliquidens, Grampus griseus, Globicephala macrorhynchus, Tursiops truncatus, and Delphinapterus leucas). The putative iron-responsive element sequence in the 5'-untranslated region of the six different dolphin species was revealed by direct sequencing of PCR fragments obtained using leukocyte genomic DNA.


RESULTS
Dolphin H and L subunits consist of 182 and 174 amino acids, respectively, and amino acid sequence identities of ferritin subunits among these dolphins are highly conserved (H: 99-100%, (99-->98) ; L: 98-100%). The conserved 28 bp IRE sequence was located -144 bp upstream from the initiation codon in the six different dolphin species.


CONCLUSIONS
These results indicate that six different dolphin species have conserved ferritin sequences, and suggest that these genes are iron-dependently expressed.
Takaesu
Azusa
A
Laboratories of Veterinary Biochemistry, School of Veterinary Medicine, Kitasato University, Aomori 034-8628, Japan. azu147@hotmail.com
Watanabe
Kiyotaka
K
Takai
Shinji
S
Sasaki
Yukako
Y
Orino
Koichi
K
D000595:Amino Acid Sequence; D000818:Animals; D001052:Apoferritins / Q000096:biosynthesis / Q000235:genetics*; D001483:Base Sequence; D004247:DNA / Q000097:blood / Q000235:genetics; D004289:Dolphins / Q000097:blood / Q000235:genetics*; D007501:Iron / Q000378:metabolism*; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D012313:RNA / Q000097:blood / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary
D012313:RNA; D004247:DNA; D001052:Apoferritins; D007501:Iron